Analysis of omega-3 fatty acid derived N-acylethanolamines in biological matrices

Renger F. Witkamp, Michiel Balvers*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)


The adequate quantification of endocannabinoids can be complex due to their low endogenous levels and structural diversity. Therefore, advanced analytical approaches, such as LC-MS, are used to measure endocannabinoids in plasma, tissues, and other matrices. Recent work has shown that endocannabinoids that are synthesized from n-3 fatty acids, such as docosahexaenoylethanolamide (DHEA) and eicosapentaenoylethanolamide (EPEA), have anti-inflammatory and anti-tumorigenic properties and stimulate synapse formation in neurites. Here, an LC-MS based method for the quantification of n-3 endocannabinoids DHEA and EPEA which is also suited to measure a wider spectrum of endocannabinoids is described. The chapter contains a step-by-step protocol for the analysis of n-3 endocannabinoids in plasma, including sample collection and solid phase extraction, LC-MS analysis, and data processing. Modifications to the protocol that allow quantifying n-3 endocannabinoids in tissues and cell culture media will also be discussed. Finally, conditions that alter endocannabinoid concentrations are briefly discussed.

Original languageEnglish
Title of host publicationEndocannabinoid signaling: Methods and Protocols
EditorsMauro Maccarrone
PublisherHumana Press Inc.
ISBN (Electronic)9781493935390
ISBN (Print)9781493935376
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Docosahexaenoylethanolamide
  • Eicosapentaenoylethanolamide
  • Endocannabinoids
  • LC-MS
  • N-3 fatty acids
  • Solid phase extraction

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